Method of using IL-11 for inflammation associated with acute pancreatitis

ABSTRACT

Provided by the present invention are methods of treating a variety of disorders including AIDS, arthritis (rheumatoid arthritis, osteoarthritis, spondyloarthropathies), antibiotic induced diarrheal diseases ( Closbidium difficile ), multiple sclerosis, osteoporosis, gingivitis, peptic ulcer disease, esophagitis, diabetes, retinitis, uveitis, reperfusion injury after myocardial infarction (MI) or cerebral vascular accident (CVA), aphthous ulcers (oral), atherosclerosis (plaque rupture), prevention of tumor metastases, asthma, preeclampsia, acute pancreatitis, psoriasis, infertility and allergic disorders such as rhinitis, conjunctivitis, and urticaria.

This application is a division of U.S. application Ser. No. 08/973,830,filed Dec. 10, 1997, now U.S. Pat. No. 5,958,401 which was the NationalStage of International Application No. PCT/US96/08059, filed May 30,1996.

FIELD OF INVENTION

The present invention relates generally to methods of treating disorderssuch as AIDS, arthritis (rheumatoid arthritis, osteoarthritis,spondyloarthropathies), antibiotic induced diarrheal diseases(Clostridium difficile), multiple sclerosis, osteoporosis, gingivitis,peptic ulcer disease, esophagitis, diabetes, retinitis, uveitis,reperfusion injury after myocardial infarction (MI), cerebral vascularaccident (CVA), aphthous ulcers (oral), atherosclerosis (plaquerupture), prevention of tumor metastases, asthma, preeclampsia, acutepancreatitis, psoriasis, infertility and allergic disorders such asrhinitis, conjunctivitis, and urticaria.

BACKGROUND OF INVENTION

Inflammatory responses include a broad range of host reaction to avariety of insults, such as injury, infection, or rejection. It is theover production of mediators that is believed to be associated with abroad range of disorders, including AIDS, arthritis (rheumatoidarthritis, osteoarthritis, spondyloarthropathies), antibiotic induceddiarrheal diseases (Clostridium difficile), multiple sclerosis,osteoporosis, gingivitis, peptic ulcer disease, esophagitis, diabetes,retinitis, uveitis, reperfusion injury after myocardial infarction (MI),cerebral vascular accident (CVA), aphthous ulcers (oral),atherosclerosis (plaque rupture), tumor metastases, asthma,preeclampsia, acute pancreatitis, psoriasis, infertility and allergicdisorders such as rhinitis, conjunctivitis, and urticaria.

These disorders and their symptoms are briefly summarized below.According to the methods of the present invention, IL-11 is administeredto modulate the hosts' over reaction to insult thereby treating thefollowing disorders.

AIDS:

Infection with HIV eventually leads to destruction of T-helper cellsproducing an immuno-compromised state. However, some immune cells, suchas the macrophage may actually be stimulated during HIV infection.HIV-infected macrophages exhibit enhanced TNF-α production when thecells are stimulated. Excessive TNF-α production has been linked toincreased pulmonary damage occurring during AIDS and has been linked tothe cachexia(weight loss) which is characteristic of the disease.

Arthritis:

Rheumatoid Arthritis:

In rheumatoid arthritis, the synovial tissue lining the joint organizesinto a mass that infiltrates and degrades articular cartilage, tendons,and bone. Normal synovial tissue consists of a thin membrane of only twoor three cell layers, comprised principally of fibroblast-like synovialcells and rare resident macrophages. In contrast, rheumatoid synovialtissue consists of a mixture of cell types: immune T- and B-cells,monocyte/macrophages, polymorphonuclear leucocytes, and thefibroblast-like cells with their rampant proliferative ability. With theexception of the fibroblasts, most of these cells are recruited to therheumatoid joint in response to inflammatory stimuli that occur as partof the pathology of this disease.

Although the etiology of rheumatoid arthritis is not clear, it issuspected that an unknown antigen, such as a bacterium, virus, ormycoplasma, is deposited in the joints as a consequence of a systemicinfection. Normally, the antigen is cleared and no disease arises;however, in genetically susceptible individuals, the antigen elicits anacute inflammatory/foreign body response in which some autologous tissuedamage occurs. This, in turn, develops into an (auto)immune response andeventually leads to a chronic inflammatory and immunologic reactionwithin the synovial lining of the joint. Thus, there is a potpourri ofactivated cell types, and the cytokines they produce continuously fuelthe proliferative and destructive ability of the synovial fibroblasts.

Osteoarthritis:

In osteoarhritis, degenerative changes to the articular cartilage,subchondral bone and the synovial membrane occur after various jointsare subjected to repeated mechanical damage. Increased levels of IL-1,TNF-α and metalloproteases have been documented within the affectedjoints of patients.

Spondyloarthropathies:

The diseases classified as spondyloarthropathy are psoriatic arthritis(PsA), juvenile chronic arthritis with late pannus onset, enterogemicspondyloartropathies (enterogenic reactive arthritis (ReA) andinflammatory bowel diseases (IBD)), urogenital spondyloarthropathies(urogenital ReA), and the undifferentiated spondyloarthropathies

In this type of arthridity, various types of immune mediated jointinflammation produce degenerative changes in the multiple joints. Thesechanges consist of inflammatory infiltration within the synovialmembranes and degenerative changes to the articular cartilage and theassociated subchondral bone. One additional feature of this particularsyndrome is that of the development of bony bridges(spondyloses) betweenthe affected joint components. Again, increased levels of IL-1, TNF-αand metalloproteases have been documented within the affected joints ofpatients.

Antibiotic Induced Diarrheal Diseases:

Yet another inflammatory disorder is antibiotic induced diarrhealdiseases (e.g., by organisms such as Clostridium difficile). Clostridiumdifficile is a common cause of diarrhea and colitis in individualsreceiving broad spectrum antibiotic therapy. Toxins released by thebacteria elicit an enterotoxigenic secretory diarrhea and also elicit anacute inflammatory response in the intestinal mucosa characterized bygranulocyte infiltration, epithelial cell necrosis, ulceration andhemorrhagic edema

Multiple Sclerosis:

Multiple sclerosis (MS) is an inflammatory demyelinating disorder of thecentral nervous system (CNS). MS is characterized histopathologically byfocal lesions in different stages of evolution in the white matter ofthe CNS. Breakdown of the blood-brain barrier and inflammatoryperivascular infiltration are the first events in lesion formation andare followed by demyelination and astrogliosis. Local inflammation ismost probably induced by an autoimmune response against the myelinsheath. Proteolytic enzymes are believed to contribute to theinflammatory tissue damage in this disease. Gelatinases, belonging tothe matrix metalloproteases, contribute to tissue destruction ininflammatory demyelinating disorders of the central nervous system suchas multiple sclerosis.

Clinical diagnosis of MS is based on the history and physical findingsindicating multiple lesions in the CNS. Although some cases areprogressive from the onset, most show remissions and exacerbations withlesions occurring in different places in the white matter. As inexperimental allergic encephalomyelitis (EAE), the symptoms of MS varyfrom one individual to another and from one time to another in anyparticular patient. EAE is a T cell-mediated autoimmune disease of theCNS.

Immune abnormalities have been described in the peripheral blood andcerebrospinal fluid of MS patients, including the presence ofinflammatory T-cells, increased synthesis of immunoregulatory cytokines,and oligoclonal immunoglobulin. Although the exact cause of MS isunknown, MS might be the consequence of auto-sensitization to myelinantigens, probably induced as cross-reactions to viral or bacterialproteins.

Osteoporosis:

Postmenopausal osteoporosis is a disorder characterized by a progressiveloss of bone tissue which begins after natural or surgical menopause andleads to the occurrence of spontaneous fractures. Although estrogendeficiency is known to cause bone loss by stimulating the resorptiveactivity of mature osteoclasts (OCs) and the proliferation anddifferentiation of OC precursors, the mechanism of these effects isstill conjectural at best. One such mechanism may be a modulatory effecton the secretion of factors that are produced in the bonemicroenvironment and influence bone remodeling. Among these are IL-1,IL-6, and tumor necrosis factor α and β (TNF). IL-1 and TNF promote boneresorption in vitro and in vivo by activating mature OCs indirectly, viaa primary effect on osteoblasts, and by stimulating the proliferationand differentiation of OC precursors. IL-6 also increases OC formationfrom hemopoietic precursors. However, IL-6 does not activate mature OCs.

Gingivitis:

Adult periodontitis is strongly associated with infection byPorphyromonas gingivalis. Proteolytic enzymes, which are produced inlarge quantity by this bacteria, are considered as important pathogenicagents. The increased production and flow of gingival crevicular fluid(GCF) is an important change in gingival tissues during periodontalinfection, correlating with clinical indices of gingival inflammation.Indeed, salivary protein and albumin concentrations of individuals withperiodontitis, which are an indication of plasma leakage due to vascularpermeability enhancement (VPE), are significantly increased compared tohealthy subjects. The production of GCF appears dependent on VPE inducedat periodontitis sites, presumably involving proteinase(s) of P.gingivalis in their generation.

Peptic Ulcer Disease:

Inhibition of gastric acid secretion with H₂-receptor antagonists and,more recently, blockers of H⁺,K⁺-ATPase (also known as the proton pump)has been the mainstay of therapy for peptic ulcer disease. Thepathophysiology of peptic ulcers remains obscure. An appreciation of thecomplexity of the physiology of the gastric mucosa has led to ahypothesis that peptic ulcers are the result of an imbalance in therelative importance of aggressive (acid, pepsin) and protective (mucus,bicarbonate, blood flow, prostaglandins, etc.) factors. Infection of themucosa of the human gastric antrum with the bacterium Helicobacterpylori has been widely accepted as the cause of chronic, active, type Bgastritis. Further, this form of gastritis has been linked directly topeptic ulcer disease by studies showing that eradication of H. pylorireverses this gastritis and prevents duodenal ulcer relapse. Becausecytokines are the principal mediators by which immune/inflammatory cellscommunicate with each other and with other cells, it is likely thatthese small peptides are involved in the pathogenesis of chronic activetype B gastritis and the resulting peptic ulcer disease.

Some cytokines (IL-1, epidermal growth factor, transforming growthfactor-α, acidic and basic fibroblast growth factors) tip the balancetowards peptic ulcer healing; others (tumor necrosis factor-α) appear tohave no effect; still others (IL-4) may even cause gastrointestinaldamage.

Esophagitis:

The most common cause of esophagitis is the chronic reflux ofhydrochloric acid from the stomach due to inefficiency of the cardiacsphincter of the stomach. The chronic presence of acid in the loweresophagus leads to damage of the esophageal mucosa In the most severeform, a syndrome called Barrett's esophagus can develop which oftenleads to esophageal cancer. Other causes of esophagitis includeparenteral chemotherapy and ionizing radiation, associated withradiation therapy for cancer in the thoracic cavity.

Diabetes:

Infiltration of the pancreatic islets by immune/inflammatory cells(insulitis), followed by loss of the insulin-producing beta cells is thecharacteristic histologic feature of insulin-dependent diabetes mellitus(IDDM).

IDDM is a T cell-mediated chronic autoimmune disease that ischaracterized by lymphocytic infiltration of the pancreatic islets ofLangerhans and by the selective destruction of insulin-producing β cellsin the islets. Since TNF-α is a proinflammatory cytokine and its gene islocalized within the major histocompatibility complex (MHC), which hasbeen shown to have a strong genetic linkage to several autoimmunedisorders including IDDM, TNF-α has been considered to be a possiblecandidate cytokine mediating the pathogenic destruction of β-isletcells.

Retinitis:

Inflammation of the light sensitive retina, retinitis, can occur due toa variety of viral, bacterial or autoimmune etiologies. The end resultis destruction of the retina and loss of sight.

Uveitis:

Inflammation of the anterior portion of the eye its associatedstructures, the iris and cornea occurs with a relatively high frequencyin patients with autoimmune disorders.

Reperfusion Injury after Myocardial Infarction (MI) and CerebralVascular Accident (CVA):

Ischemia-induced endothelial cell injury has been described as thepivotal causative event leading to an array of pathophysiologic sequelaesuch as microvascular vasoconstriction, adhesion and aggregation ofplatelets and neutrophils, and deceased blood flow, inclusivelydescribed as the “no reflow-phenomenon” early after reperfusion.

The infiltration and activation of multiple types of inflammatory cellsresult in a series of degenerative changes in the vasculature of theaffected area, as well as inciting damage of the surrounding parenchymaltissue.

Aphthous Ulcers (Oral):

Although the cause of aphthous ulcers remain unknown, many physiciansbelieve they are caused by autoimmune phenomena, which cause thedestruction of discrete areas of the oral mucosa which leads to oralulceration. Among the cytokines present in these active areas ofulceration, TNF-α appears to play a predominant role.

Age-related diseases such as atherosclerosis (plaque rupture), fibrosis,osteoporosis, and many others, are associated with increased levels ofcertain cytokines such as IL-1, IL-6 and TNF-α, suggesting thatphysiological aging in humans is associated with an increased capabilityof peripheral blood mononuclear cells to produce proinflammatorycytokines. Although atherosclerosis contributes to the narrowing of theblood vessel lumen, often the large atherosclerotic plaques cause fewerproblems than smaller plaques. This occurs due to the sudden rupture ofmedium sized plaques. It appears that this rupture is associated withincreased concentrations of various metalloproteases, probably derivedfrom inflammatory cells within the plaque.

Prevention of Tumor Metastases:

The processes of tumor invasion and metastasis are thought to dependupon increased proteolytic activity of invading tumor cells. Matrixmetalloproteinases, cathepsins B, D, and L, and plasminogen activatorhave been proposed to participate in the metastatic cascade. Cathepsin Dhas been suggested to be an independent marker of prognosis in breastcancer.

Acute Pancreatitis:

Acute pancreatitis is severe inflammation of the pancreas which oftenresults in pancreatic necrosis. Death resulting from acute pancreatitisensues as a result of multiple organ failure and other clinical symptomsresembling sepsis. In the early stages of acute pancreatitis, elevatedserum levels of IL-1, IL-6 and TNF-α are frequently seen.

Psoriasis:

Psoriasis is a chronic inflammatory skin disorder involvinghyperproliferation of the epidermis and inflammation of both theepidermis and the dermis. Macrophage and neutrophil infiltration of thedermis and epidermis is seen, and proinflanmatory mediators are releasedfrom these activated cells.

Infertility:

Infertility frequently involves disorders of the ovary which result inabnormal folliculogenesis and inflammatory reactions. Leukocytes arefound in the follicular fluid, which, when activated can produce IL-1,IL-6 and TNF-α. Accordingly, IL-11 is administered to promotefolliculogenesis, oocyte maturation, ovulation and early embryogenesiswithin the oviduct.

Asthma:

Asthma is a disease with two major components, a marked inflammatoryreaction and a disorder of bronchial smooth muscle reactivity producingbronchospasm. Increased production of inflammatory mediators causesinfiltration of leukocytes, with lymphocytes, eosinophils and mast cellsbeing present in large quantities.

Allergic Disorders such as:

Rhinitis, Conjunctivitis and Urticaria (“Hives”):

In these three types of inflammation, multiple allergens are capable ofevoking the infiltration and activation of “allergic” classes ofleukocytes, i.e., eosinophils, mast cells and basophils, resulting inthe subsequent release of histamine, platelet activating factor, etc.Given this general type of reaction, these aforementioned disordersoccur in the nose, conjunctiva and the skin.

Preeclampsia:

Preeclampsia is characterized by development of hypertension,endothelial cell disruption, coagulopathy, leukocyte activation, edema,renal dysfunction, and fetal growth disturbances after the twentiethweek of pregnancy and occurs in 8% to 10% of all pregnancies in theUnited States annually. The endothelial cell damage seen in preeclampsiamay be produced in part by TNF-α.

It has been recently reported that IL-11 is produced by placentalfibroblasts and that it causes proliferation and differentiation of thetrophoblast, the major cell type of the placenta. In non-pregnantindividuals, IL-11 causes a physiologic plasma volume expansion which isvery similar to that caused by normal pregnancy. IL-11 also appears tobe able to decrease TNF-α by activated macrophages. In preeclampsia,trophoblast growth and differentiation are abnormal, plasma volumeexpansion fails to occur and TNF-α levels are elevated, indirectlysuggesting a deficiency of IL-11.

In Graft vs. Host Disease (GVHD), the immunologic recognition andresponse seen are believed to be caused by histocompatibilitydifferences between the donor and recipient and cytotoxicity byalloreactive T cells. The cellular injury in GVHD is thought to becaused by cellular infiltration of effector cells into target tissueswith resultant destruction. The theory of the destruction seen in GVHDwas based on the observation of lymphocytes juxtaposed to dying cells(satellitosis) observed frequently in the skin of patients or animalswith GVHD. Pulmonary complications are often lethal components of acuteGVHD.

BRIEF SUMMARY OF THE INVENTION

Provided by the present invention are methods of treating a variety ofdisorders including AIDS, arthritis (rheumatoid arthritis,osteoarthritis, spondyloarthropathies), antibiotic induced diarrhealdiseases (Clostridium difficile), multiple sclerosis, osteoporosis,gingivitis, peptic ulcer disease, esophagitis, diabetes, retinitis,uveitis, reperfusion injury after myocardial infarction (MI) or cerebralvascular accident (CVA), aphthous ulcers (oral), atherosclerosis (plaquerupture), prevention of tumor metastases, asthma, preeclampsia, acutepancreatitis, psoriasis, infertility and allergic disorders such asrhinitis, conjunctivitis, and urticaria.

According to the present invention, IL-11, analogs, and derivativesthereof, are administered to patients, either prophylactically or at theonset of symptoms associated with the aforementioned disorders. IL-11can be administered in suitable pharmaceutically acceptable carrierseither alone or in combination with other conventional agents useful inalleviating the symptoms associated with the aforementioned disorders. Asuitable IL-11 formulation comprises, for example, 5 mg IL-11, 3.10 mghistidine, and 22.5 mg glycine, for example, as a lyophilized powderwhich can be reconstituted with 1 ml sterile water for injection. IL-11can be supplied intravenously or can be applied topically informulations to the nasal mucosa or the conjunctiva or oral mucosa as anaqueous drop formulation or mouthwash, respectively. In localizedsurface reactions, a topical formulation is preferred whereas in moresevere generalized body-wide inflammatory states, parental routes, suchas subcutaneous or intravenous injection are preferred. Suitable dosesof IL-11 are in the range of 1-50 μg/kg subcutaneous for multiple dailydoses and for shorter periods of treatment in severe inflammatorystates, doses are increased to 50 to 100 μg/kg subcutaneous orintravenous. Doses are administered daily for between one day and sixmonths, or for as long as is deemed necessary and safe in the treatmentof the aforementioned disorders, as is readily ascertained by standardtests by the attending physician, depending upon the nature of thedisorder being treated.

DETAILED DESCRIPTION OF THE INVENTION

Provided by the present invention are methods for using IL-11 for thetreatment of AIDS, arthritis (rheumatoid arthritis, osteoarthritis,spondyloarthropathies), antibiotic induced diarrheal diseases(Clostridium difficile), multiple sclerosis, osteoporosis, gingivitis,peptic ulcer disease, esophagitis, diabetes, retinitis, uveitis,reperfusion injury after myocardial infarction (MI), cerebral vascularaccident (CVA), aphthous ulcers (oral), atherosclerosis (plaquerupture), prevention of tumor metastases, asthma, preeclampsia, acutepancreatitis, psoriasis, infertility and allergic disorders such asrhinitis, conjunctivitis, and urticaria

Interleukin 11 (IL-11) is a pleiotropic cytokine that stimulatesprimitive lymphohematopoietic progenitor cells and synergizes with otherhematopoietic growth factors to stimulate the proliferation andmaturation of megakaryocytes. IL-11 is described in detail inInternational Application PCT/US90/06803, published May 30, 1991; aswell as in U.S. Pat. No. 5,215,895; issued Jun. 1, 1993. A cloned humanIL-11 was previously deposited with the ATCC, 10801 UniversityBoulevard, Manassa, Va. 20110-2209, on Mar. 30, 1990 under ATCC No.68284. Moreover, as described in U.S. Pat. No. 5,270,181; issued Dec.14, 1993; and U.S. Pat. No. 5,292,646; issued Mar. 8, 1994; IL-11 mayalso be produced recombinantly as a fusion protein with another protein.IL-11 can be produced in a variety of host cells by resort to nowconventional genetic engineering techniques. In addition, IL-11 can beobtained from various cell lines, for example, the human lung fibroblastcell line, MRC-5 (ATCC Accession No. CCL 171) and Paul et al., the humantrophoblastic cell line, TPA30-1 (ATCC Accession No. CRL 1583).Described in Proc Natl Acad Sci USA 87:7512 (1990) is a cDNA encodinghuman IL-11 as well as the deduced amino acid sequence (amino acids 1 to199). U.S. Pat. No. 5,292,646, supra, describes a des-Pro form of IL-11in which the N-terminal proline of the mature form of IL-11 (amino acids22-199) has been removed (amino acids 23-199). As is appreciated by oneskilled in the art, any form of IL-11, which retains IL-11 activity, isuseful according to the present invention.

In addition to recombinant techniques, IL-11 may also be produced byknown conventional chemical synthesis. Methods for constructing thepolypeptides useful in the present invention by synthetic means areknown to those of skill in the art. The synthetically constructedcytokine polypeptide sequences, by virtue of sharing primary, secondary,or tertiary structural and conformational characteristics with thenatural cytokine polypeptides are anticipated to possess biologicalactivities in common therewith. Such synthetically constructed cytokinepolypeptide sequences or fragments thereof, which duplicate or partiallyduplicate the functionality thereof may also be used in the method ofthis invention. Thus, they may be employed as biologically active orimmunological substitutes for the natural, purified cytokines useful inthe present invention.

Modifications in the protein, peptide or DNA sequences of thesecytokines or active fragments thereof may also produce proteins whichmay be employed in the methods of this invention. Such modifiedcytokines can be made by one skilled in the art using known techniques.Modifications of interest in the cytokine sequences, e.g., the IL-11sequence, may include the replacement, insertion or deletion of one ormore selected amino acid residues in the coding sequences. Mutagenictechniques for such replacement, insertion or deletion are well known toone skilled in the art. (See, e.g., U.S. Pat. No. 4,518,584.)

Other specific mutations of the sequences of the cytokine polypeptideswhich may be useful therapeutically as described herein may involve,e.g., the insertion of one or more glycosylation sites. Anasparagine-linked glycosylation recognition site can be inserted intothe sequence by the deletion, substitution or addition of amino acidsinto the peptide sequence or nucleotides into the DNA sequence. Suchchanges may be made at any site of the molecule that is modified byaddition of O-linked carbohydrate. Expression of such altered nucleotideor peptide sequences produces variants which may be glycosylated atthose sites.

Additional analogs and derivatives of the sequence of the selectedcytokine which would be expected to retain or prolong its activity inwhole or in part, and which are expected to be useful in the presentmethod, may also be easily made by one of skill in the art. One suchmodification may be the attachment of polyethylene glycol (PEG) ontoexisting lysine residues in the cytokine sequence or the insertion ofone or more lysine residues or other amino acid residues that can reactwith PEG or PEG derivatives into the sequence by conventional techniquesto enable the attachment of PEG moieties.

Additional analogs of these selected cytokines may also be characterizedby allelic variations in the DNA sequences encoding them, or inducedvariations in the DNA sequences encoding them. It is anticipated thatall analogs disclosed in the above-referenced publications, includingthose characterized by DNA sequences capable of hybridizing to thedisclosed cytokine sequences under stringent hybridization conditions ornon-stringent conditions (Sambrook et al, Molecular Cloning. ALaboratory Manual, 2d edit, Cold Spring Harbor Laboratory, New York(1989)) will be similarly useful in this invention.

Also considered useful in these methods are fusion molecules, preparedby fusing the sequence or a biologically active fragment of the sequenceof one cytokine to another cytokine or proteinaceous therapeutic agent,e.g., IL-11 fused to IL-6 (see, e.g., methods for fusion described inPCT/US91/06186 (WO92/04455), published Mar. 19, 1992). Alternatively,combinations of the cytokines may be administered together according tothe method.

Thus, where in the description of the methods of this invention IL-11 ismentioned by name, it is understood by those of skill in the art thatIL-11 encompasses the protein produced by the sequences presentlydisclosed in the art, as well as proteins characterized by themodifications described above yet which retain substantially similaractivity in. Standard laboratory tests are utilized to monitor progressof the treatment Levels of TNF-α in serum or the biologic effects ofTNF-α could be followed in a variety of these diseases. Decreasedsymptomatology could also be used to monitor the effectiveness oftreatment as is well known to physicians skilled in the art of treatingsuch disorders.

The present invention thus involves treating patients having disorderssuch as AIDS, arthritis (rheumatoid arthritis, osteoarthritis,spondyloardiropathies), antibiotic induced diarrheal diseases(Clostridium difficile), multiple sclerosis, osteoporosis, gingivitis,peptic ulcer disease, esophagitis, diabetes, retinitis, uveitis,reperfusion injury after myocardial infarction (MI), cerebral vascularaccident (CVA), aphthous ulcers (oral), atherosclerosis (plaquerupture), prevention of tumor metastases, asthma, preeclampsia, acutepancreatitis, psoriasis, infertility and allergic disorders such asrhinitis, conjunctivitis, and urticaria and involves administering aneffective amount of IL-11 in a pharmaceutical carrier. Treatment ispreferably prophylactic, but may also be at the onset of symptomsassociated with the aforementioned disorders.

Suitable pharmaceutically acceptable carriers facilitate administrationof IL-11 and are well known in the art. Exemplary carriers includesterile saline, lactose, sucrose, calcium phosphate, gelatin, dextrin,agar, pectin, peanut oil, olive oil, sesame oil, and water.Additionally, the carrier or diluent includes a time delay material,such as glyceryl monostearate or glyceryl distearate alone or with awax. In addition, slow release polymer formulations can be used.Suitable sustain-release matrices contain the active ingredient in amixture with one or more of the following: sodium bentonite,ethylcellulose, stearic acid, calcium stearate, adipic acid, fumaricacid, polyethylene glycol, deacetylated chitin, and cellulose acetate.Suitable preservatives and/or stabilizers may be included.

Alternatively, IL-11 can be combined with other conventional agentsuseful in alleviating the symptoms associated with the aforementioneddisorders, as is readily apparent to one skilled in the art.

A suitable IL-11 formulation comprises, for example, 5 mg IL-11, 3.10 mghistidine and 22.5 mg glycine, e.g., as a lyophilized powder which canbe reconstituted with 1 mL sterile water for injection. As is apparentto one skilled in the art, other suitable formulations are equallyeffective according to the methods of the present invention. Moreover,the IL-11 can be applied topically in formulations to the nasal mucosaor the conjunctiva or oral mucosa as an aqueous drop formulation ormouthwash, respectively. In localized surface reactions, the topicalformulation could be used, while in more severe generalized body-wideinflammatory states, the parenteral routes could be employed,subcutaneous or intravenous injection. Suitable doses of IL-11 may be inthe range of 1-50 μg/kg SC for multiple daily doses, but for shorterperiods of treatment in severe inflammatory states doses may beincreased to 50-100 μg/kg SC or IV. In the treatment of theaforementioned disorders, IL-11 can be administered by any suitableroute, but certain routes are preferable for certain disorders, e.g.,administered systemically, i.e., parenterally. Of the parenteral routes,subcutaneous and intravenous are preferred.

A suitable treatment regimen for patients undergoing treatment,including for example prophylactic treatment, may be determined by theattending physician based upon such factors as the patient's age, sex,weight, and general health. Generally, a suitable dose of cytokine,e.g., IL-11 ranges broadly, preferably between 1 and 100 μg/kg bodyweight. Another suitable dose may be in the range of about 10 to 50μg/kg and about 50 μg/kg with a preferable amount of 25 μg IL-11 perkilogram of body weight. If desired, these doses can be adjusted tounits. A unit is conventionally described as the concentration ofpolypeptide which leads to half-maximal stimulation in a suitable assay,e.g., for IL-11, the T1165 assay described in PCT/US90/06803. Doses maybe administered daily for between one day and six months, or for as longas is deemed necessary and safe, as is readily ascertained by standardtests by the attending physician, depending upon the nature of thedisorder being treated. Where appropriate, the dosages may be adjustedupward or downward, for example, a dosing regimen requiringadministration of IL-11 at a dose of 25 μg/kg, daily for one week, orfewer days, or multiple weeks if indicated. The progress of treatment isappropriately monitored by measurement of markers associated with thedisorder being treated to determine if such a dose results in a decreaseof for example, INF-α levels (or corresponding marker) and if not,increasing the dose two-fold for an additional time period of treatmentand measurement of marker levels until an effective dosing regimen isreached.

The following examples illustrate the methods of the present inventionand in particular the use of IL-11 in treating AIDS, arthritis(rheumatoid arthritis, osteoarthitis, spondyloarthropathies), antibioticinduced diarrheal diseases (Clostridium difficile), multiple sclerosis,osteoporosis, gingivitis, peptic ulcer disease, esophagitis, diabetes,retinitis, uveitis, reperfusion injury after myocardial infarction (MI),cerebral vascular accident (CVA), aphthous ulcers (oral),atherosclerosis (plaque rupture), prevention of tumor metastases,asthma, acute pancreatitis, psoriasis, infertility and allergicdisorders such as rhinitis, conjunctivitis, and urticaria. However, theexamples do not limit the scope of the invention in any way.

Example 1 describes the treatment of arthritis (rheumatoid arthritis,osteoarthritis, spondyloarthropathies). Example 2 relates to thetreatment of antibiotic induced diarrheal diseases (Clostridiumdifficile); Example 3 describes the treatment of multiple sclerosis;Example 4 relates to the treatment of osteoporosis; Example 5 describesthe treatment of peptic ulcer disease; and Example 6 relates to thetreatment of infertility.

EXAMPLE 1

Treatment of Arthritis (Rheumatoid Arthritis, Osteoarthritis,Spondyloarthropathies)

HLA-B27 transgenic Fischer 344 rats spontaneously exhibit lesions of thegastrointestinal system, the joints, the skin and the gonads whichappear similar to the spondylarthropies in humans that have beenassociated with the HLA-B27 and β₂-microglobulin genes (Hammer et al.,Cell 63:1099(1990)). Overt inflammatory bowel disease is observed in100% of the rats by 20 weeks of age. Recently, Scofield et al., PNAS90:9330 (1993) reported that short portions of the primary amino acidsequence of the hypervariable regions of HLA-B27 share homology with theproteins of gram-negative bacteria and that they are capable of bindingthe HLA-B27 molecule. After birth as the gastrointestinal tract iscolonized with bacteria, tolerance to the HLA-B27 molecule is broken asthe mucosa is exposed to the luminal bacteria This results in thedevelopment of an autoimmune phenomenon which leads to a chronicinflammatory syndrome manifest initially in the gastrointestinal tract.

During studies of these rats, administration of rhIL-11 reduced theswelling and redness of tarsal and knee joints of these animals. Blindedassessment of radiographs of the joints of these animals revealed that 4of 6 vehicle treated animals had radiographic evidence of jointdistension and osteophyte formation, while only one of 6 rhIL-11 treatedanimal had radiographic changes. Subsequent histologic evaluationsconfirmed the presence of pannus formation, degeneration of thearticular cartilage and osteophyte formation in the vehicle treatedanimals. rhIL-11 treated animals exhibited minimal to no lesions.

EXAMPLE 2

Treatment of Antibiotic Induced Diarrheal Diseases (Clostridiumdifficile)

Toxin A from C. difficile, binds to a brush border toxin receptor,causing secretion of fluid, increased intestinal permeability and anintense inflammatory infiltrate in rat ileum. As shown below,pretreatment with rhIL-11 reduces the intestinal effects of toxin A.

Adult rats were injected (SC) with either rhIL-11 (150 μg/kg) or vehicle20 min before administration of toxin A. Rat ileal loops (5 cm) werethen injected with toxin A (5 μg) or buffer (0.4 ml) and 4 hours later,enterotoxicity was assessed by fluid secretion (mg/cm) andblood-to-lumen excretion of ³H-mannitol (dpm/loop).

TABLE 1 Results Secretion Permeability Buffer (n = 8) 109 + 6  1,450 +270 Toxin A (n = 12) 388 + 14** 45,200 + 4,500** rhIL-11 + Toxin A (n =5) 211 + 15**++  5,460 + 2,030*++ n = number of rats tested, *p < 0.05and **p < 0.01 vs Buffer, ++p < 0.01 vs Toxin A

Histologic evaluation indicated that rhIL-11 attenuated epithelialdamage caused by toxin A, but had no effect on neutrophil infiltrationor congestion and edema of the mucosa Preincubation of rhIL-11 (50μg/ml) with purified toxin A (5 μg/ml) in vitro did not alter toxinA-induced cell rounding in cultured lung fibroblasts or ³H-toxin Aspecific binding to rat ileal brush borders.

rhIL-11 significantly inhibits C difficile toxin A-mediated secretion(by 45.6%) and permeability (by 88%) and reduces mucosal damage in vivo,but has no apparent effect on fibroblast rounding or toxin A binding toits membrane receptor in vitro.

EXAMPLE 3

Treatment of Multiple Sclerosis

Experimental autoimmune encephalomyelitis (EAE) is considered the bestavailable animal counterpart for multiple sclerosis (MS) and has thusbeen widely used to study potential immunoregulatory mechanisms involvedin the pathogenesis of this disease. As the T cell subsets, cytokines,and cellular adhesion molecules that mediate successful diseaseinduction have been extensively characterized, the model serves as avaluable tool for delineating the biological actions of immunoregulatoryproteins. In susceptible strains of mice, disease can be induced byinjection of encephalitogenic proteins (myelin basic protein (MBP) orproteolipid protein (PLP)) in complete Freund's adjuvant (CFA).Alternatively, EAE can be passively transferred to naive animals withMBP or PLP-sensitized T lymphocytes, encephalitogenic T cell lines, or Tcell clones. The clinical course of disease is characterized by weightloss and a progressive paralysis which commonly leads to completebilateral hind limb paralysis. The paralytic episode coincides with anacute perivascular inflammatory response in the central nervous system(CNS) which is comprised predominantly of infiltrating macrophages and Tcells. In most species the disease remits spontaneously with thesequence of recovery being the reverse of that of onset.

Based on the pivotal role for macrophage derived cytokines in EAE andthe ability of IL-11 to inhibit RAW cell derived TNF-α production invitro, the effects of IL-11 administration are evaluated in an adoptivetransfer model of EAE. The protocol for adoptive transfer is describedin detail previously in Leonard, J. Exp. Med. 181:381 (1995). Briefly,spleen or lymph node cells from PLP immunized mice are cultured in vitrowith antigen for 96 hours and subsequently transferred to naive mice.30×10⁶ PLP stimulated spleen or lymph node cells are used in eachexperiment. Clinical signs of EAE are graded as follows: 0.5, distallimp tail; 1, complete limp tail; 1.5. limp tail with unsteady gait; 2.0partial hind limb paralysis; 3.0 complete hind limb paralysis.

Administration of IL-11 (250 μg/kg/day as single SC injections) for thefirst 10 days following PLP stimulated spleen or lymph node celltransfer dramatically altered the course of EAE with both the incidence,as well as the severity of disease reduced by IL-11 treatment. The meanscore for controls, (n=10) is greater than 2.5 whereas for IL-11-treated(n=5), the mean score is less than half of the control.

To determine if IL-11 influences T cell activation/expansion in vitro,LNC from PLP primed mice were stimulated with antigen and IL-11 (500ng/ml) prior to cell transfer. The addition of IL-11 during antigenstimulation had no effect on the ability of the cells to transferdisease to naive mice.

EXAMPLE 4

Treatment of Osteoporosis

Following estrogen depletion, osteoporosis occurs secondary to increasedlytic activity of osteoclasts. The effects of rhIL-11 on murineosteoporosis induced by estrogen depletion were assessed in C57BlackMice following ovariectomy. Animals were ovariectomized, and two weekslater rhIL-11 therapy was commenced at 250 μg/kg SC BID for 7 days. Theanimals were killed and histologic assessment of osteoclast activity inthe tibia was performed using TRAP staining. rhIL-11 decreased TRAPstaining as compared to saline treated animals(OVX+saline 5.76±2.63%versus OVX+rhIL-11 2.93±1.24, p<0.0001). Thus, the IL-11 treated animalshad fewer osteoclasts and therefore, less potential for bone loss.

EXAMPLE 5

Treatment of Peptic Ulcer Disease

The effects of three different dosages of recombinant humaninterleukin-11 (rhIL-11), given subcutaneously (SC) either prior to orsubsequent to intracolonic administration of trinitrobenzene sulfonicacid (TNB), were studied in Sprague-Dawley rats. The TNB or control weregiven in a 40% ethanol solution to 312 anesthetized adult male ratsallotted to one of 26 groups (n=12). Control groups were: subcutaneous(SC); saline alone; intrarectal (IR); 40% ethanol alone; TNB alone; 40%ethanol alone, and SC, rhIL-11 at the highest dosage alone and groupscombining TNB with rhIL-11 therapy, testing three dosages (100, 300, and1,000 μg/kg), given either before or after induction of colitis withTNB. Body weight changes were monitored. Rats were euthanized at 3 days,7 days, or 14 days after TNB administration. At necropsy, samples werecollected to evaluate fecal occult blood, mucosal myeloperoxidaseactivity and mucosal gross indexes of ulceration. Histopathologic andultrastructural analyses of the colonic mucosa were performed. The TNBalone elicited a prolonged, severe colitis in treated animals, and theethanol control group showed a short-lasting, less severe colonicinflammatory response. Colonic ulcer indexes of rhIL-11 treated ratsshowed a consistent, dose-related reduction in the severity of theTNB-induced colitis, whether the interleukin was given before or afterthe TNB. This reduction was significant (P<0.05) after administration ofthe intermediate (300 μg/kg) and highest (1,000 μg/kg) dose levels ofrhIL-11, in the groups given rhIL-11 for 7 days after TNB.Myeloperoxidase activity was increased during the TNB-induced colitisand was reduced by rhIL-11 administration (P<0.01). Fecal occult bloodloss increased with colitis and paralleled its severity. rhIL-11enhanced mucus production and decreased the severity of TNB-inducedcolitis. Thus, given these beneficial effects of IL-11 in this colitismodel, IL-11 can be used to ameliorate peptic ulcer disease.

EXAMPLE 6

Treatment of Infertility

Follicular fluid (FF) represents the medium which ensures oocytematuration, fertilization, transport, and facilitates in vitro spermcapacitation. Leukocytes are found in the follicular fluid, which, whenactivated can produce IL-1, IL-6 and TNF-α.

The periovulatory FF is assayed and is found to contain detectablequantities of IL-11. IL-11 was assayed and found to be present in sixfrom seven samples of pooled follicular fluid (85.7%), in variableamounts (19.776-70.546 pg/ml). None of the 13 serum samples containedIL-11. IL-11 was measurable in 67.2% of the follicular fluids whichcontained a normally fertilized oocyte, 66.7% of the immature follicles,38.5% of the postmature follicles, and 84.2% of the atretic follicles.The amount of IL-11 was significantly higher in atretic follicles, whencompared to all the other groups (p<0.05). These data are evidence of abiphasic peak in IL-11 during the normal events of folliculardevelopment, IL-11 is present during early follicle development, fallsas the follicle becomes post mature, and increases again during atresia.Considering the significantly higher level of IL-11 in atreticfollicles, IL-11 is involved in follicular atresia which occurs in 99%of all normal follicles that initially develop. Accordingly, IL-11 isadministered to promote folliculogenesis, oocyte maturation, ovulation,normal atresia of unovulated follicles, and early embryogenesis withinthe oviduct

While the present invention has been described in terms of specificmethods and compositions, it is understood that variations andmodifications will occur to those skilled in the art upon considerationof the present invention.

Numerous modifications and variations in the invention as described inthe above illustrative examples are expected to occur to those skilledin the art and, consequently, only such limitations as appear in theappended claims should be placed thereon. Accordingly, it is intended inthe appended claims to cover all such equivalent variations which comewithin the scope of the invention as claimed.

What is claimed:
 1. A method of treating inflammation associated withacute pancreatitis, comprising administering to a patient in needthereof a pharmaceutically effective amount of IL-11 in an amountsufficient to reduce said inflammation.
 2. The method of claim 1,wherein the amount of IL-11 is between 1 and 100 μg/kg body weight. 3.The method of claim 1, wherein the amount of IL-11 is between 1 and 50μg/kg body weight.
 4. The method of claim 1, wherein the IL-11 isadministered prophylactically to said patient.
 5. The method of claim 1,wherein said IL-11 is administered at the onset of symptoms of acutepancreatitis in said patient.
 6. The method of claim 1, wherein saidIL-11 is administered parenterally to said patient.
 7. The method ofclaim 1, wherein said IL-11 is administered intravenously orsubcutaneously to said patient.
 8. The method of claim 1, whereinadministration of IL-11 to said patients decreases TNF-α levels in saidpatient.
 9. The method of claim 1, wherein said IL-11 is provided in aformulation comprising histidine.
 10. The method of claim 1, whereinsaid IL-11 is provided in a formulation comprising glycine.
 11. Themethod of claim 1, wherein said acute pancreatitis is early stage acutepancreatitis.
 12. The method of claim 1, wherein said IL-11 isrecombinantly produced.